Concentrating proteins from solution - (Mar/08/2004 )
I am trying to concentrate few proteins that are present in a culture medium (with lot of salts and carbohydrates) in a nanomolar concentrations. Can any one suggest a method to concentrate the proteins to load on a gel?
I appreciate it very much
very easy, http://www.millipore.com/ should have the products you want
It depends on what resources you are working with and how rich your lab is. If Millipore products are not your lab chief's idea of good budgeting, there are other ways. Grow the culture in 100 ml batches. Overgrow it unless you have a limit on how much growth should happen because cells tend to lyse in late stationary. Centrifuge the cells down. Dialyze 25 ml against a suitable buffer with 50% glycerol. This will bring down your volume to 6 ml. You can either electrophorese and do a silver stain or plan on an FITC conjugation to increase sensitivity to picomolar range.
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